I am trying to conjugate a protein in a MWCNT (Multi-Walled Carbon Nanotube) to investigate the improvement of protein citotoxicity. But It's not showing clear effect and not sure if the nanotube is not improving the protein activity or if it does not having conjugation between the protein and the nanotube. Is there any technique that can evaluate the conjugation to discard one of the hypotheses?
Dear Caio,
You can try FTIR spectral analysis (before and after adding protein into CNTs)...
FTIR is a great idea, so long as the product can be successfully purified (IR doesn't like impure samples).
I'd be tempted to either run a size exclusion column or an electrophoretic gel. The conjugation product should be significantly large than the two starting species.
I tried to make a electrophoresis to compare the band of protein and conjugate. However, the nanotube is positively charged and so does not carry the race in the electric field. Perhaps the best alternative is the IR. Thank you all!
Caio, in that case, you could run electrophoresis the "old fashioned way".
Make up a gel in a horizontal trough, and cut a hole/channel in the centre for your sample using a DNA/agarose set up (you may need to trim the area of the gel containing the loading wells). You would then run the gel as normal, and your unbound proteins would run one way, the CNT-bound protein the other.
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